Inducible T cell receptors and uses thereof

Abstract

Combinations comprising one or more nucleic acid molecules comprising a nucleic acid sequence A encoding for a TCR alpha chain linked to a dimerization domain, and a nucleic acid sequence B encoding for a TCR beta chain linked to a dimerization domain, as well as proteins encoded by such nucleic acid molecules and corresponding uses and methods.

The invention claimed is: 
     
       1. A combination comprising one or more nucleic acid molecules, said one or more nucleic acid molecules comprising:
 (a) a nucleic acid sequence A, comprising
 (i) a nucleic acid sequence encoding an amino acid sequence being at least 90% identical to SEQ ID NO: 2 and optionally comprising at least one amino acid substitution compared to the amino acid sequence of SEQ ID NO: 2 selected from the group consisting of T44A and T47A, and 
 (ii) a nucleic acid sequence encoding an inducible dimerization domain linked to, and downstream of, said nucleic acid sequence of (a)(i); and 
 
 (b) a nucleic acid sequence B, comprising
 (i) a nucleic acid sequence encoding an amino acid sequence being at least 90% identical to SEQ ID NO: 4 and comprising at least one amino acid substitution compared to the amino acid sequence of SEQ ID NO: 4 selected from the group consisting of K4V, N5P, Y37K, L63A, S77A and R79A, and 
 (ii) a nucleic acid sequence encoding an inducible dimerization domain linked to, and downstream of, said nucleic acid of (b)(ii), wherein said dimerization domain corresponds to the dimerization domain of (a)(ii), 
 
 wherein said amino acid sequence of (a)(i) comprises at least one amino acid substitution compared to SEQ ID NO: 2. 
 
     
     
       2. The combination of  claim 1 , wherein the nucleic acid sequence A and the nucleic acid sequence B are comprised by separate nucleic acid molecules or comprised in one nucleic acid molecule. 
     
     
       3. The combination of  claim 1 , wherein
 (c) said nucleic acid sequence of (a)(i) encodes an amino acid sequence comprising two amino acid substitutions compared to the amino acid sequence of SEQ ID NO: 2, said two substitutions being T44A and T47A; and 
 (d) said nucleic acid sequence of (b)(i) encodes an amino acid sequence comprising three amino acid substitutions compared to the amino acid sequence of SEQ ID NO: 4, wherein said three substitutions are K4V, N5P and Y37K, or said three substitutions are L63A, S77A, and R79A. 
 
     
     
       4. The combination of  claim 1 , wherein the dimerization domain is a homodimerization domain or a heterodimerization domain optionally selected from the group consisting of ERT2, FKBP, CalcineurinA (CNA), CyP-Fas, GyrB, GAI, GID1, eDHFR and FRB domain of mTOR. 
     
     
       5. An expression cassette system comprising:
 the combination of  claim 1  as a single unitary expression cassette or 
 the combination of  claim 1  as first and second expression cassettes, wherein the first expression cassette comprises the nucleic acid sequence A and the second expression cassette comprises the nucleic acid sequence B. 
 
     
     
       6. A vector comprising the expression cassette system of  claim 5 . 
     
     
       7. The vector of  claim 6 , wherein the vector is a retroviral vector or a lentiviral vector. 
     
     
       8. A kit comprising:
 the combination of  claim 1 , and 
 a dimerization agent corresponding to the dimerization domain encoded by the nucleic acid sequence (a)(ii) and the dimerization domain encoded by the nucleic acid sequence (b)(ii), said dimerization agent being capable of inducing dimerization of said dimerization domains. 
 
     
     
       9. The kit of  claim 8 , wherein the dimerization agent is selected from the group consisting of 4-hydroxytamoxifen, Endoxifen, 4-(1-[4-(Dimethylaminoethoxy) phenyl]-2-phenyl-1-butenyl) phenol, AP21967, 23,27-Epoxy-3H-pyrido [2,1-c][1,4]oxaazacyclohentriacontine, FK1012, FK506, FKCsA, Rapamycin, Coumermycin, Gibberellin. 
     
     
       10. A host cell comprising the combination of  claim 1 . 
     
     
       11. The host cell of  claim 10 , wherein the host cell is a T lymphocyte. 
     
     
       12. The host cell of  claim 10 , wherein said host cell comprises a dimerization agent corresponding to the dimerization domain encoded by the nucleic acid sequence (a) (ii) and the dimerization domain encoded by the nucleic acid sequence (b)(ii), said dimerization agent being capable of inducing dimerization of said dimerization domains. 
     
     
       13. The host cell of  claim 12 , wherein the dimerization agent is selected from the group consisting of 4-hydroxytamoxifen, Endoxifen, 4-(1-[4-(Dimethylaminoethoxy) phenyl]-2-phenyl-1-butenyl) phenol, AP21967, 23,27-Epoxy-3H-pyrido [2,1-c][1,4]oxaazacyclohentriacontine, FK1012, FK506, FKCsA, Rapamycin, Coumermycin, Gibberellin. 
     
     
       14. A protein encoded by the combination of  claim 1 . 
     
     
       15. A method for dimerizing a protein encoded by the combination of  claim 1 , comprising:
 combining a population of proteins encoded by nucleic acid sequence A and nucleic acid sequence B with a dimerization agent corresponding to the dimerization domain encoded by the nucleic acid sequence (a)(ii) and the dimerization domain encoded by the nucleic acid sequence (b)(ii), said dimerization agent being capable of inducing dimerization of said dimerization domains to said population of proteins. 
 
     
     
       16. A method for preparing an inducible T cell receptor, comprising:
 introducing the combination of  claim 1  in vitro into a host cell under conditions allowing the expression of the protein encoded by nucleic acid sequence A and nucleic acid sequence B. 
 
     
     
       17. A method for the treatment of cancer, comprising administering the combination as defined in  claim 1 , the expression cassette system as defined in  claim 5 , the vector as defined in  claim 6 , the host cell as defined in  claim 10 , or the protein as defined in  claim 8  to a subject in need thereof. 
     
     
       18. The method of  claim 17 , wherein said cancer is solid cancer or blood cancer. 
     
     
       19. A pharmaceutical composition comprising:
 the combination as defined in  claim 1 , the expression cassette system as defined in  claim 6 , the vector as defined in  claim 6 , the host cell as defined in  claim 10 , or the protein as defined in  claim 14 ; and 
 a pharmaceutically acceptable excipient.